Study Liver Cytochrome P450 3A4 Inhibition and Hepatotoxicity Using DMSO-Differentiated HuH-7 Cells

  • Yitong LiuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1473)


Metabolically competent, inexpensive, and robust in vitro cell models are needed for studying liver drug-metabolizing enzymes and hepatotoxicity. Human hepatoma HuH-7 cells develop into a differentiated in vitro model resembling primary human hepatocytes after a 2-week dimethyl sulfoxide (DMSO) treatment. DMSO-treated HuH-7 cells express elevated cytochrome P450 3A4 (CYP3A4) enzyme gene expression and activity compared to untreated HuH-7 cells. This cell model could be used to study CYP3A4 inhibition by reversible and time-dependent inhibitors, including drugs, food-related substances, and environmental chemicals. The DMSO-treated HuH-7 model is also a suitable tool for investigating hepatotoxicity. This chapter describes a detailed methodology for developing DMSO-treated HuH-7 cells, which are subsequently used for CYP3A4 inhibition and hepatotoxicity studies.

Key words

HuH-7 DMSO CYP3A4 Inhibition Hepatotoxicity 


  1. 1.
    Lin J, Schyschka L, Muhl-Benninghaus R, Neumann J, Hao L, Nussler N, Dooley S, Liu L, Stockle U, Nussler AK, Ehnert S (2012) Comparative analysis of phase I and II enzyme activities in 5 hepatic cell lines identifies Huh-7 and HCC-T cells with the highest potential to study drug metabolism. Arch Toxicol 86(1):87–95. doi: 10.1007/s00204-011-0733-y CrossRefPubMedGoogle Scholar
  2. 2.
    Donato MT, Jover R, Gomez-Lechon MJ (2013) Hepatic cell lines for drug hepatotoxicity testing: limitations and strategies to upgrade their metabolic competence by gene engineering. Curr Drug Metab 14(9):946–968CrossRefPubMedGoogle Scholar
  3. 3.
    Nakabayashi H, Taketa K, Miyano K, Yamane T, Sato J (1982) Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Res 42(9):3858–3863PubMedGoogle Scholar
  4. 4.
    Choi S, Sainz B Jr, Corcoran P, Uprichard S, Jeong H (2009) Characterization of increased drug metabolism activity in dimethyl sulfoxide (DMSO)-treated Huh7 hepatoma cells. Xenobiotica 39(3):205–217. doi: 10.1080/00498250802613620 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Liu Y, Flynn TJ, Xia M, Wiesenfeld PL, Ferguson MS (2015) Evaluation of CYP3A4 inhibition and hepatotoxicity using DMSO-treated human hepatoma HuH-7 cells. Cell Biol Toxicol 31(4-5):221–230. doi: 10.1007/s10565-015-9306-9 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Hisaka A, Ohno Y, Yamamoto T, Suzuki H (2010) Prediction of pharmacokinetic drug-drug interaction caused by changes in cytochrome P450 activity using in vivo information. Pharmacol Ther 125(2):230–248. doi: 10.1016/j.pharmthera.2009.10.011 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Division of Applied ToxicologyOffice of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug AdministrationLaurelUSA

Personalised recommendations